The Influence Of Polyporus Polysaccharides On The Mouse Bone Marrow-derived Dendritic Cell And Its Anti-hepatoma H22 Tumor Effect

Background and ObjectivePolyporus polysaccharides(PPS) extracted from polyporus umbellatus Fries, which is a kind of traditional Chinese medicine.The studies have shown that PPS can improve the body's immunopotency and have effect against tumor.Dendritic cells(DC) are the most important professional antigen-presenting cells (APC) known so far.As the initiator of primary immune response,DC can capture, process,present antigen,and play critical roles in anti-tumor immune regulation.The membrane surfaces of mature dendritic cells not only express high level of major histocompatibility complex(MHC) molecules,but also accessory and adhesion molecules.They can effectively present antigens to MHCⅠand MHCⅡmolecules, and at the same microenvironment make help T cells and kill T cells tussocks so that promote cytotoxic T Lymphocyte(CTL) response.The previous studies indicate that DC function decreases in tumor patients.And the quantities of tumor infiltrating dendritic cell increase in tumor microenvironment that are positive correlation with tumor patients' prognosis.Whether immune pharmacology of the traditional Chinese medicine is related to tumor-bearing body's DC that influences their multiplication,maturation,phenotype and changes of function? The researches of anti-tumor immunity pharmacological mechanism about traditional Chinese medicine on DC provide a new perspective for the anti-tumor immunology.In this study,we observed the influences of PPS on the multiplication, maturation,surface mark,antigen presentation of DC which derived from the bone marrow in H22-bearing mice.And we surveyed immune effect about PPS for H22-bearing mice.We discussed the anti-tumor effect and mechanism of PPS,and provided the experiment basis for further explicating immunocompetence of PPS.Methods1.The experimental H22 hepatoma bearing mice were made.2.Influences of PPS on DC in vitro:The H22 hepatoma bearing mice were killed after being feed ten days,for obtaining the bone marrow cells under aseptic condition,synchronously using the GM-CSF and IL-4 to induce them into dendritic cells.While dendritic cells were cultured with different concentrations(1μg/ml, 5μg/ml,10μg/ml) of PPS,and another group with normal salineCNS) as a control group.And at the 5th day,H22 cell lysate made by freeze-thaw method were added into dendritic cells,continuously incubated until the 8th day.The appearance character of DC was observed by microscope.Dendritic cells were collected 8 days later,and then the expressions of CD11a and CD86 were analyzed by flow cytometry (FCM).The cytotoxicity effect of DC that active T cells on H22 hepatoma cells was examined with MTT.As the same way,dendritic cells were cultured but not added H22 cell lysate made by freeze-thaw method,collected 8 days later and analyzed DC cell cycle.3.Effects of PPS on the H22 hepatoma bearing mice:tumor-bearing KM mice were made,and 24 hours later,the different concentrations of PPS(20μg,200μg, 2000μg) were injected into the left abdominal cavities of mice,while the equal volume of NS ones as the comparative group.Lasting 10 days,the last 24 hours later, these mice were killed.Then,immune effect of PPS was observed by counting spleen index and thymus index of mice,and the anti-tumor effect of PPS was examined with inhibition rate of tumor.All the data were expressed as mean value±S.D.,and analyzed by statistical software SPSS10.0.One-way analysis of variance was used during the groups. P-value＜0.05 was considered to be statistically significant.Result1.After adding H22 cell lysate made by freeze-thaw method,PPS could improve the maturation of bone marrow-derived dendritic cells from the mice,and the expressions of CD86 and CD11a were increased in PPS groups(compared with the control group,there was significant difference,p＜0.05).The killing rate of DC activated CTL was also increased in PPS groups(compared with the control group, there was significant difference,p＜0.05).Moreover,PPS up-regulated proliferation of DC which not added H22 cell lysate made by freeze-thaw method(compared with the control group,there was significant difference,p＜0.05).2.Compared with control group,thc spleen index of the H22 hepatoma bearing mice were increased in PPS groups(p＜0.05).But the thymus index was not obviously increased(p＞0.05).Moreover,the PPS improved the anti-tumor effect of the H22 hepatoma bearing mice that inhibited the growth of tumor(p＜0.05).Conclusions1.PPS can promote DC derived from bone marrow in H22-bearing mice proliferation.2.PPS and H22 cell lysate made by freeze-thaw method can synergistically improve the differentiation and maturation of DC derived from bone marrow in H22-bearing mice and can increase the expressions of CD11a and CD86,promote ability of CTL.3.PPS has positive effect on mice immune function through inhibiting the growth of tumor.